ABSTRACT
Chlamydia trachomatis (CT) is the leading cause of bacterial sexually transmitted diseases worldwide, which can cause diseases such as pelvic inflammatory disease, and cervical and fallopian tube inflammation, and poses a threat to human health. Rosmarinic acid (RosA) is an active ingredient of natural products with anti-inflammatory and immunomodulatory effects. This study aimed to investigate the role of RosA in inhibiting autophagy-regulated immune cells-CD8+ T cells via the Ras/Raf/MEK/ERK signaling pathway in a CT-infected mouse model. Mice were inoculated with CT infection solution vaginally, and the mechanistic basis of RosA treatment was established using H&E staining, flow cytometry, immunofluorescence, transmission electron microscopy, and western blot. The key factors involved in RosA treatment were further validated using the MEK inhibitor cobimetinib. Experimental results showed that both RosA and the reference drug azithromycin could attenuate the pathological damage to the endometrium caused by CT infection; flow cytometry showed that peripheral blood CD8+ T cells increased after CT infection and decreased after treatment with RosA and the positive drug azithromycin (positive control); immunofluorescence showed that endometrial CD8 and LC3 increased after CT infection and decreased after RosA and positive drug treatment; the results of transmission electron microscopy showed that RosA and the positive drug azithromycin inhibited the accumulation of autophagosomes; western bolt experiments confirmed the activation of autophagy proteins LC3â ¡/â , ATG5, Beclin-1, and p62 after CT infection, as well as the inhibition of Ras/Raf/MEK/ERK signaling. RosA and azithromycin inhibition of autophagy proteins activates Ras/Raf/MEK/ERK signaling. In addition, the MEK inhibitor cobimetinib attenuated RosA's protective effect on endometrium by further activating CD8+ T cells on a CT-induced basis, while transmission electron microscopy, immunofluorescence, and western blots showed that cobimetinib blocked ERK signals activation and further induced phagocytosis on a CT-induced basis. These data indicated that RosA can activate the Ras/Raf/MEK/ERK signaling pathway to inhibit autophagy, and RosA could also regulate the activation of immune cells-CD8+T cells to protect the reproductive tract of CT-infected mice.
Subject(s)
Autophagy , CD8-Positive T-Lymphocytes , Chlamydia Infections , Chlamydia trachomatis , Cinnamates , Depsides , MAP Kinase Signaling System , Rosmarinic Acid , Animals , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/drug effects , Autophagy/drug effects , Female , Chlamydia Infections/immunology , Chlamydia Infections/drug therapy , Chlamydia trachomatis/drug effects , Chlamydia trachomatis/immunology , Mice , Depsides/pharmacology , MAP Kinase Signaling System/drug effects , Cinnamates/pharmacology , ras Proteins/metabolism , raf Kinases/metabolism , Disease Models, Animal , Mice, Inbred C57BLABSTRACT
Objective To investigate the effect of doxycycline on the proliferation of human small cell lung cancer H446 cells in vitro and its possible mechanisms. Methods H446 cells were treated with different concentrations of doxycycline for 24,48,72 or 96 h. Cell proliferation was measured by CCK-8 assay,IC50 was caculated by Bliss,and cell apoptosis was examined by TUNEL assay. The protein and mRNA expression of Bcl-2 and Bax in H446 cells were detected by Western blot and real time-PCR,respectively. Re-sults Doxycycline inhibited the proliferation of H446 cells in a concentration-time dependent manner. The half inhibitory concentra-tions(IC50)of doxycycline on cell proliferation in H446 cells were approximately 24.10 mg/L for 24 h treatment group,10.98 mg/L for 48 h treatment group,8.20 mg/L for 72 h treatment group and 8.03 mg/L for 96 h treatment group,respectively. Furthermore,doxycy-cline treatment could also induce the apoptosis of H446 cells in a concentration-dependent manner. Moreover,doxycycline treatment dramatically down-regulated the expression of Bcl-2 and up-regulated the expression of Bax in H446 cells both at the protein and at the mRNA levels. The differences were statistically significant(P<0.05). Conclusion Doxycycline can inhibit the proliferation of H446 cells by inducing apoptosis via down-regulating the expression of Bcl-2 and up-regulating the expression of Bax. Doxycycline has great potential as a chemotherapeutic agent for human small cell lung cancer.
ABSTRACT
Objective By selecting the prescriptions of treating blood group incompatibility in modern literature, and exploring their regularity, to establish the overall thought of traditional Chinese medicine in treatment.Methods 90 articles on maternal-fetal blood group incompatibility were found.Statistical methods were adopted to summarize high frequently used medicines and formula. Results High frequently used medicines were Virgate Wormwood Herb, Baical Skullcap Root, Rhubarb, Cape Jasmine Fruit, White peony Alba, Chinese Angelica,Largehead Atractylodes Rhizome,Motherwort Herb,and Milkvetch Root, which were mentioned for 238 times in the literatures, occupying 72.79% of the total frequency. Conclusion Based on this result,heat-clearing and dampness-removing, activating blood circulation to eliminate blood stasis was setup as the principle for treating this disease.
ABSTRACT
0.05).CONCLUSION:Domestic octreotide acetate have same effect to imported octreotide acetate in the treatment of Esophageal and gastric Varices Bleeding which is caused by portal hypertension in patients with decompensated liver cirrhosis.Stopping bleeding rate of the achieved the similar level with.
